Research Article
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Year 2018, Volume: 3 Issue: 3, 300 - 310, 10.10.2018
https://doi.org/10.28978/nesciences.468960

Abstract

References

  • AOAC, 1996. Official Methods of Analyses. Assoc. Official Analytical Chemists, Arlington, VA.
  • Armstrong, R.N. (1997). Structure, catalytic mechanism, and evolution of the glutathione transferases. Chemical Research in Toxicology, 10, 2-18.
  • Atamer, A., Bilici, A., Yenice, N., Selek, S., İlhan, N. & Atamer, Y. (2008). The importance of paraoxonase 1 activity, nitric oxide and lipid peroxidation in hepatosteatosis. The Journal of International Medical Research, 36, 771-776.
  • Aviram, M., Billecke, S., Sorenson, R., Bisgaier, C., Newton, R., Rosenblat, M., Erogul, J., Hsu, C. & Dunlop, C. (1998). Paraoxonase Active Site required for protection against LDL oxidation involves its free sulfhydryl group and is different from that required for its arylesterase/paraoxonase activities. Arteriosclerosis, Thrombosis, and Vascular Biology, 18, 1617-1624.
  • Aviram, M. & Rosenblat, M. (2004). Paraoxonases 1, 2, and 3, Oxidative stress, and macrophage foam cell formation during atherosclerosis development. Free Radical Biology and Medicine, 37(9), 1304-1316.
  • Baldissera, M.D., Souza, C.F., Doleski, P.H., Monteiro, S.G., da Silva, A.S. & Baldisserotto, B. (2018). Serum adenosine deaminase and xanthine oxidase activities in silver catfish naturally infected with Ichthyophthirius multifiliis: The influence of these enzymes on inflammatory and oxidative status. Journal of Fish Diseases, 41, 263-268.
  • Bastos, V.L.F.C., Folly, E., Rossini, A., Ceccarelli, P.S., Senhorini, J.A. & Bastos, J.C. (1998). Paraoxonase activity in liver of pacu, Piaractus mesopotamicus Holmberg (Characidae). Revista Brasileira de Zoologia, 15(5), 677-685.
  • Bodolay, E., Seres, I., Szodoray, P., Csipo, I., Jakab, Z., Vegh, J., et al. (2008). Evaluation of paraoxonase activity in patients with mixed connective tissue disease. Journal of Rheumatology, 35, 237-43.
  • Camps, J., Marsillach, J. & Joven, J. (2009). Measurement Of serum paraoxonase-1 activity in the evaluation of liver function. World Journal of Gastroenterology, 15(16), 1929-1933.
  • Carver, J.D. & Walker, W.A. (1995). The role of nucleotides in human nutrition. Nutrition Biochemistry, 6, 58-72.
  • Craig, S.R. & McLean, E. (2005). The organic movement: a role for NuPro® as an alterative protein source. In: Nutritional Biotechnology in the Food and Feed Industry, K.A. Jacques & T.P. Lyons, (Eds). Nottingham University Press, UK.
  • Deng, D., Mei, C., Mai, K., Tan, B. P., Ai, Q., & Ma, H. (2013). Effects of a yeast-based additive on growth and immune responses of white shrimp, Litopenaeus vannamei (Boone, 1931), and aquaculture environment. Aquaculture Research, 44, 1348-1357.
  • Fegan, D.E. (2006). Functional foods for aquaculture: benefits of Nupro and dietary nucleotides in aquaculture feeds. In: Nutritional Biotechnology in the Feed and food Industries. Proceedings of Alltech’s 22nd Annual Symposium, T.P. Lyons, K.A. Jacques & J.M.Hower (Eds). Pub. Nottingham University press.
  • Ferreira, I., Pinho, O., Vieira, E., Tavarela, J.G. (2010). Brewer's Saccharomyces yeast biomass: characteristics and potential applications. Trends in Food Science & Technology, 21, 77-84.
  • Ganguly, S., Paul. I., Kumar, S. & Mukhopadhayay, S.K. (2010). Application and effectiveness of immunostimulants, probiotics, and prebiotics in aquaculture: A Review. The Israeli Journal of Aquaculture-Bamidgeh, 62(3), 130-138.
  • Giusti, G. & Galanti. B. (1984). Colorimetric method. In: H.U. Bergmeyer, (Eds). Methods of Enzymatic Analysis.1st edition. Weinheim, Germany: Verlag Chemie.
  • Habig, W.H., Pabst, M.J. & Jakoby, W.B. (1974). Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry, 249, 7130-7139.
  • Hasko, G. & Cronstein, B. N. 2004. Adenosine: an endogenous regulator of innate immunity. Trends Immunol, 25, 33-39.
  • Hayes, J.D., Flanagan, J.U. & Jowsey, I.R. (2005). Glutathione transferases. Annual review of pharmacology and toxicology, 45, 51-88.
  • Keleştemur, G.T. & Özdemir, Y. (2011). Balıklarda Antioksidan Savunma ve Oksidatif Stres. Türk Bilimsel Derlemeler Dergisi, 4(1), 69-73.
  • Li, P. & Gatlin, III D.M. (2003). Evaluation of brewers’ yeast. (Saccharomyces cereviciae) as a feed supplement for hybrid striped bass (Marone chrysops x M. saxatillis). Aquaculture, 219, 681-692.
  • Li, W.F., Costa, L.G. & Furlong, C.E. (1993). Serum paraoxonase status: a major factor in determining resistance to organophosphates. Journal of Toxicology and Environmental Health, 40, 337-346.
  • Li, P. & Gatlin III D.M. (2006). Nucleotide nutrition in fish: Current knowledge and future applications. Aquaculture, 251, 141-152.
  • Lin, Y.H., Wang, H. & Shiau, S. (2009). Dietary nucleotide supplementation enhances growth and immune responses of grouper, Epinephelus malabaricus. Aquaculture Nutrition, 15, 117-122.
  • Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193, 265-275.
  • Mani, R., Meena, B., Valivittan, K. & Suresh, A. (2014). Glutathione –S-Transferase and catalase activity in different tissues of marine catfish Arius arius on exposure to cadmium. International Journal of Pharmacy and Pharmaceutical Sciences, 6(1), 326-332.
  • Manoppo, H., Manurung, U.S. & Tumbol, R.A. (2015). Efficacy of baker’s yeast as immunostimulant in Nile tilapia (Oreochromis niloticus) International Journal of ChemTech Research, 8(3), 1396-1402.
  • Martinez-Alvarez, R.M., Morales, A.E. & Sanz, A. (2005). Antioxidant defenses in fish: Biotic and abiotic factors. Reviews in Fish Biology and Fisheries, 15, 75-88.
  • Meng, Y., Ma, R., Ma, J., Han, D., Xu, W., Zhang, W. & Mai, K. (2017). Dietary nucleotides improve the growth performance, antioxidative capacity and intestinal morphology of turbot (Scophthalmus maximus). Aquaculture Nutrition, 23, 585-593.
  • Moreno, E., Canet, J., Gracia, E., Liuis, C., Mallol, J., Canela, EI., Cortes, A. & Casado, V. (2018). Molecular evidence of adenosine deaminase linking adenosine A2A receptor and CD26 proteins. Front Pharmacol, 9, 106, 18 p.
  • National Research Council (NRC), (1993). Nutrient requirements of fish. National Academy Press, Washington DC.
  • Özlüer-Hunt, A., Özkan-Yılmaz, F., Berköz, M., Engin, K., Gündüz, SG. & Yalın, S. (2016). Effects of Dietary Nucleotide Yeast on Immune Responses and Antioxidant Enzyme Activities of Rainbow Trout Juveniles (Oncorhynchus mykiss) The Israeli Journal of Aquaculture - Bamidgeh, 68, 1264, 12 pp.
  • Peng, M., Xu, W., Ai, Q., Mai, K., Liufu Z. & Zhang, K. (2013). Effects of nucleotide supplementation on growth, immune responses and intestinal morphology in juvenile turbot fed diets with graded levels of soybean meal (Scophthalmus maximus L.). Aquaculture, 392-395, 51-58.
  • Repetto, M., Semprine, J., & Boveris, A. (2012). Lipid peroxidation: chemical mechanism, biological implications and analytical determination. In: Lipid peroxidation, A. Catala (Ed) New Delhi, Intech, 1-28.
  • Ringø, E., Olsen, RE., Gonzales-Vecino, JL., Wadsworth, S. & Song, SK. (2012) Use of immunostimulants and nucleotides in aquaculture: A review. Journal of Marine Science: Research and Development, 2, 22 pp.
  • Sanchez-Pozo, A. & Gil, A. (2002). Nucleotides as semiessential nutritional components. British Journal of Nutrition, 87, 135-137.
  • Sauer, N., Mosenthin, R. & Bauer, E. (2011). The role of dietary nucleotides in single-stomached animals. Nutrition Research Reviews, (2011), 24, 46-59.
  • Shiau, S., Gabaudan J. & Lin, Y. (2015). Dietary nucleotide supplementation enhances immune responses and survival to Streptococcus iniae in hybrid tilapia fed diet containing low fish meal. Aquaculture Reports, 2, 77-81.
  • Smith R.R. (1989). Nutritional Energetics. In: J.E. Halver, (Ed) Fish Nutrition, 2. Edn. Academic Press, San Diego, C, pp 476-549. Sugimoto, M. (1995). Glutathione S-transferases (GSTs). Nippon Rinsho, 53, 1253-1259.
  • Uauy, R., Quan, R. & Gil, A. (1994) Role of nucleotides in intestinal development and repair: implications for infant nutrition. Journal of Nutrition, 124, 1436-1441.
  • Walsh, M.J., Tsao, K.L., & Leleiko, N.S., (1992). Characterization of DNA-protein interactions within a distal regulatory element upstream of a mammalian housekeeping gene promoter. Journal of Biological Chemistry, 267, 7026-7035.
  • Welker, T.L., Lim, C., Yildirim-Aksoy, M., & Klesius, P.H. (2011). Effects of dietary supplementation of a purified nucleotide mixture on immune function and disease and stress resistance in channel catfish, Ictalurus punctatus. Aquaculture Research, 42, 1878-1889.
  • Whitehead, J., Wadsworth, S. & Carr, I. (2006). The power of purified nucleotides. Aquac Health Int, 4, 14-16.
  • Xiang X, Zhou X, Chen J, & Zheng Z. 2011. Effects of yeast nucleotide on growth performance, body composition and immune indices of common carp (Cyprinus carpio). Chinese Journal of Animal Nutrition, 23, 171.
  • Xiong, J., Jin, M., Yuan, Y., Luo, JX., Lu, Y., Zhou, QC., Liang, C., &Tan, ZL. 2018. Dietary nucleotide-rich yeast supplementation improves growth, innate immunity and intestinal morphology of Pacific white shrimp (Litopenaeus vannamei). Aquaculture Nutrition, 1-11.
  • Xu L, Ran C, He SX, Zhang JL, Hu J, Yang YL, et al. (2015). Effects of dietary yeast nucleotides on growth, non-specific immunity, intestine growth and intestinal microbiota of juvenile hybrid tilapia Oreochromis niloticus ♀ × Oreochromis aureus ♂. Animal Nutrition, 1: 244-51.
  • Yuan, X. Y., Liu, W. B., Liang, C., Sun, C. X., Xue, Y. F., Wan, Z. D. & Jiang, G. Z. (2017). Effects of partial replacement of fish meal by yeast hydrolysate on complement system and stress resistance in juvenile Jian carp (Cyprinus carpio var. Jian). Fish & Shellfish Immunology, 67, 312–321.

Investigation of Glutathion S-Transferase, Adenosine deaminase, Paraoxonase Activities in Liver of Oncorhynchus mykiss Fed with Nucleotide-Yeast Supplemented Diet

Year 2018, Volume: 3 Issue: 3, 300 - 310, 10.10.2018
https://doi.org/10.28978/nesciences.468960

Abstract

In this study, it was evaluated Glutathione S transferase (GST),
Adenosine deaminase (ADA), and paraoxonase (PON) activities in liver tissue of
Oncorhynchus
mykiss
fed with nucleotide yeast base protein supplemented diet. Throughout
the 60-day period the control group was fed a fish meal based basal diet, and
three other groups were fed diets in which 20% (NP 20), 40% (NP 40) and 60% (NP
60) fish meal was substituted with nucleotide (Nu-Pro® (NP) yeast). At the end
of experiment, liver tissue GST, PON and ADA activity was increased
significantly (P<0.05) in nucleotide-yeast groups when compared to control
group. 

References

  • AOAC, 1996. Official Methods of Analyses. Assoc. Official Analytical Chemists, Arlington, VA.
  • Armstrong, R.N. (1997). Structure, catalytic mechanism, and evolution of the glutathione transferases. Chemical Research in Toxicology, 10, 2-18.
  • Atamer, A., Bilici, A., Yenice, N., Selek, S., İlhan, N. & Atamer, Y. (2008). The importance of paraoxonase 1 activity, nitric oxide and lipid peroxidation in hepatosteatosis. The Journal of International Medical Research, 36, 771-776.
  • Aviram, M., Billecke, S., Sorenson, R., Bisgaier, C., Newton, R., Rosenblat, M., Erogul, J., Hsu, C. & Dunlop, C. (1998). Paraoxonase Active Site required for protection against LDL oxidation involves its free sulfhydryl group and is different from that required for its arylesterase/paraoxonase activities. Arteriosclerosis, Thrombosis, and Vascular Biology, 18, 1617-1624.
  • Aviram, M. & Rosenblat, M. (2004). Paraoxonases 1, 2, and 3, Oxidative stress, and macrophage foam cell formation during atherosclerosis development. Free Radical Biology and Medicine, 37(9), 1304-1316.
  • Baldissera, M.D., Souza, C.F., Doleski, P.H., Monteiro, S.G., da Silva, A.S. & Baldisserotto, B. (2018). Serum adenosine deaminase and xanthine oxidase activities in silver catfish naturally infected with Ichthyophthirius multifiliis: The influence of these enzymes on inflammatory and oxidative status. Journal of Fish Diseases, 41, 263-268.
  • Bastos, V.L.F.C., Folly, E., Rossini, A., Ceccarelli, P.S., Senhorini, J.A. & Bastos, J.C. (1998). Paraoxonase activity in liver of pacu, Piaractus mesopotamicus Holmberg (Characidae). Revista Brasileira de Zoologia, 15(5), 677-685.
  • Bodolay, E., Seres, I., Szodoray, P., Csipo, I., Jakab, Z., Vegh, J., et al. (2008). Evaluation of paraoxonase activity in patients with mixed connective tissue disease. Journal of Rheumatology, 35, 237-43.
  • Camps, J., Marsillach, J. & Joven, J. (2009). Measurement Of serum paraoxonase-1 activity in the evaluation of liver function. World Journal of Gastroenterology, 15(16), 1929-1933.
  • Carver, J.D. & Walker, W.A. (1995). The role of nucleotides in human nutrition. Nutrition Biochemistry, 6, 58-72.
  • Craig, S.R. & McLean, E. (2005). The organic movement: a role for NuPro® as an alterative protein source. In: Nutritional Biotechnology in the Food and Feed Industry, K.A. Jacques & T.P. Lyons, (Eds). Nottingham University Press, UK.
  • Deng, D., Mei, C., Mai, K., Tan, B. P., Ai, Q., & Ma, H. (2013). Effects of a yeast-based additive on growth and immune responses of white shrimp, Litopenaeus vannamei (Boone, 1931), and aquaculture environment. Aquaculture Research, 44, 1348-1357.
  • Fegan, D.E. (2006). Functional foods for aquaculture: benefits of Nupro and dietary nucleotides in aquaculture feeds. In: Nutritional Biotechnology in the Feed and food Industries. Proceedings of Alltech’s 22nd Annual Symposium, T.P. Lyons, K.A. Jacques & J.M.Hower (Eds). Pub. Nottingham University press.
  • Ferreira, I., Pinho, O., Vieira, E., Tavarela, J.G. (2010). Brewer's Saccharomyces yeast biomass: characteristics and potential applications. Trends in Food Science & Technology, 21, 77-84.
  • Ganguly, S., Paul. I., Kumar, S. & Mukhopadhayay, S.K. (2010). Application and effectiveness of immunostimulants, probiotics, and prebiotics in aquaculture: A Review. The Israeli Journal of Aquaculture-Bamidgeh, 62(3), 130-138.
  • Giusti, G. & Galanti. B. (1984). Colorimetric method. In: H.U. Bergmeyer, (Eds). Methods of Enzymatic Analysis.1st edition. Weinheim, Germany: Verlag Chemie.
  • Habig, W.H., Pabst, M.J. & Jakoby, W.B. (1974). Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry, 249, 7130-7139.
  • Hasko, G. & Cronstein, B. N. 2004. Adenosine: an endogenous regulator of innate immunity. Trends Immunol, 25, 33-39.
  • Hayes, J.D., Flanagan, J.U. & Jowsey, I.R. (2005). Glutathione transferases. Annual review of pharmacology and toxicology, 45, 51-88.
  • Keleştemur, G.T. & Özdemir, Y. (2011). Balıklarda Antioksidan Savunma ve Oksidatif Stres. Türk Bilimsel Derlemeler Dergisi, 4(1), 69-73.
  • Li, P. & Gatlin, III D.M. (2003). Evaluation of brewers’ yeast. (Saccharomyces cereviciae) as a feed supplement for hybrid striped bass (Marone chrysops x M. saxatillis). Aquaculture, 219, 681-692.
  • Li, W.F., Costa, L.G. & Furlong, C.E. (1993). Serum paraoxonase status: a major factor in determining resistance to organophosphates. Journal of Toxicology and Environmental Health, 40, 337-346.
  • Li, P. & Gatlin III D.M. (2006). Nucleotide nutrition in fish: Current knowledge and future applications. Aquaculture, 251, 141-152.
  • Lin, Y.H., Wang, H. & Shiau, S. (2009). Dietary nucleotide supplementation enhances growth and immune responses of grouper, Epinephelus malabaricus. Aquaculture Nutrition, 15, 117-122.
  • Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193, 265-275.
  • Mani, R., Meena, B., Valivittan, K. & Suresh, A. (2014). Glutathione –S-Transferase and catalase activity in different tissues of marine catfish Arius arius on exposure to cadmium. International Journal of Pharmacy and Pharmaceutical Sciences, 6(1), 326-332.
  • Manoppo, H., Manurung, U.S. & Tumbol, R.A. (2015). Efficacy of baker’s yeast as immunostimulant in Nile tilapia (Oreochromis niloticus) International Journal of ChemTech Research, 8(3), 1396-1402.
  • Martinez-Alvarez, R.M., Morales, A.E. & Sanz, A. (2005). Antioxidant defenses in fish: Biotic and abiotic factors. Reviews in Fish Biology and Fisheries, 15, 75-88.
  • Meng, Y., Ma, R., Ma, J., Han, D., Xu, W., Zhang, W. & Mai, K. (2017). Dietary nucleotides improve the growth performance, antioxidative capacity and intestinal morphology of turbot (Scophthalmus maximus). Aquaculture Nutrition, 23, 585-593.
  • Moreno, E., Canet, J., Gracia, E., Liuis, C., Mallol, J., Canela, EI., Cortes, A. & Casado, V. (2018). Molecular evidence of adenosine deaminase linking adenosine A2A receptor and CD26 proteins. Front Pharmacol, 9, 106, 18 p.
  • National Research Council (NRC), (1993). Nutrient requirements of fish. National Academy Press, Washington DC.
  • Özlüer-Hunt, A., Özkan-Yılmaz, F., Berköz, M., Engin, K., Gündüz, SG. & Yalın, S. (2016). Effects of Dietary Nucleotide Yeast on Immune Responses and Antioxidant Enzyme Activities of Rainbow Trout Juveniles (Oncorhynchus mykiss) The Israeli Journal of Aquaculture - Bamidgeh, 68, 1264, 12 pp.
  • Peng, M., Xu, W., Ai, Q., Mai, K., Liufu Z. & Zhang, K. (2013). Effects of nucleotide supplementation on growth, immune responses and intestinal morphology in juvenile turbot fed diets with graded levels of soybean meal (Scophthalmus maximus L.). Aquaculture, 392-395, 51-58.
  • Repetto, M., Semprine, J., & Boveris, A. (2012). Lipid peroxidation: chemical mechanism, biological implications and analytical determination. In: Lipid peroxidation, A. Catala (Ed) New Delhi, Intech, 1-28.
  • Ringø, E., Olsen, RE., Gonzales-Vecino, JL., Wadsworth, S. & Song, SK. (2012) Use of immunostimulants and nucleotides in aquaculture: A review. Journal of Marine Science: Research and Development, 2, 22 pp.
  • Sanchez-Pozo, A. & Gil, A. (2002). Nucleotides as semiessential nutritional components. British Journal of Nutrition, 87, 135-137.
  • Sauer, N., Mosenthin, R. & Bauer, E. (2011). The role of dietary nucleotides in single-stomached animals. Nutrition Research Reviews, (2011), 24, 46-59.
  • Shiau, S., Gabaudan J. & Lin, Y. (2015). Dietary nucleotide supplementation enhances immune responses and survival to Streptococcus iniae in hybrid tilapia fed diet containing low fish meal. Aquaculture Reports, 2, 77-81.
  • Smith R.R. (1989). Nutritional Energetics. In: J.E. Halver, (Ed) Fish Nutrition, 2. Edn. Academic Press, San Diego, C, pp 476-549. Sugimoto, M. (1995). Glutathione S-transferases (GSTs). Nippon Rinsho, 53, 1253-1259.
  • Uauy, R., Quan, R. & Gil, A. (1994) Role of nucleotides in intestinal development and repair: implications for infant nutrition. Journal of Nutrition, 124, 1436-1441.
  • Walsh, M.J., Tsao, K.L., & Leleiko, N.S., (1992). Characterization of DNA-protein interactions within a distal regulatory element upstream of a mammalian housekeeping gene promoter. Journal of Biological Chemistry, 267, 7026-7035.
  • Welker, T.L., Lim, C., Yildirim-Aksoy, M., & Klesius, P.H. (2011). Effects of dietary supplementation of a purified nucleotide mixture on immune function and disease and stress resistance in channel catfish, Ictalurus punctatus. Aquaculture Research, 42, 1878-1889.
  • Whitehead, J., Wadsworth, S. & Carr, I. (2006). The power of purified nucleotides. Aquac Health Int, 4, 14-16.
  • Xiang X, Zhou X, Chen J, & Zheng Z. 2011. Effects of yeast nucleotide on growth performance, body composition and immune indices of common carp (Cyprinus carpio). Chinese Journal of Animal Nutrition, 23, 171.
  • Xiong, J., Jin, M., Yuan, Y., Luo, JX., Lu, Y., Zhou, QC., Liang, C., &Tan, ZL. 2018. Dietary nucleotide-rich yeast supplementation improves growth, innate immunity and intestinal morphology of Pacific white shrimp (Litopenaeus vannamei). Aquaculture Nutrition, 1-11.
  • Xu L, Ran C, He SX, Zhang JL, Hu J, Yang YL, et al. (2015). Effects of dietary yeast nucleotides on growth, non-specific immunity, intestine growth and intestinal microbiota of juvenile hybrid tilapia Oreochromis niloticus ♀ × Oreochromis aureus ♂. Animal Nutrition, 1: 244-51.
  • Yuan, X. Y., Liu, W. B., Liang, C., Sun, C. X., Xue, Y. F., Wan, Z. D. & Jiang, G. Z. (2017). Effects of partial replacement of fish meal by yeast hydrolysate on complement system and stress resistance in juvenile Jian carp (Cyprinus carpio var. Jian). Fish & Shellfish Immunology, 67, 312–321.
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Details

Primary Language English
Subjects Hydrobiology
Journal Section 3
Authors

Ferbal Özkan-yılmaz

Arzu Özlüer-hunt This is me

Mehmet Berköz This is me

Metin Yıldırım This is me

Publication Date October 10, 2018
Submission Date May 28, 2018
Published in Issue Year 2018 Volume: 3 Issue: 3

Cite

APA Özkan-yılmaz, F., Özlüer-hunt, A., Berköz, M., Yıldırım, M. (2018). Investigation of Glutathion S-Transferase, Adenosine deaminase, Paraoxonase Activities in Liver of Oncorhynchus mykiss Fed with Nucleotide-Yeast Supplemented Diet. Natural and Engineering Sciences, 3(3), 300-310. https://doi.org/10.28978/nesciences.468960

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